Trypanosoma caninum n. sp. (Protozoa: Kinetoplastida) isolated from intact skin of a domestic dog ( Canis familiaris) captured in Rio de Janeiro, Brazil.

An unknown Trypanosoma species was isolated from an axenic culture of intact skin from a domestic dog captured in Rio de Janeiro, Brazil, which was co-infected with Leishmania (Viannia) braziliensis. Giemsa-stained smears of cultures grown in different media revealed the presence of epimastigotes, trypomastigotes, spheromastigotes, transitional stages, and dividing forms (epimastigotes or spheromastigotes). The highest frequency of trypomastigotes was observed in RPMI (15.2%) and DMEM (9.2%) media containing 5% FCS, with a mean length of these forms of 43.0 and 36.0 mum, respectively. Molecular analysis by sequential application of PCR assays indicated that this trypanosome differs from Trypanosoma cruzi and T. rangeli when specific primers were applied. On the other hand, a PCR strategy targeted to the D7 domain of 24salpha rDNA, using primers D75/D76, amplified products of about 250 bp in that isolate (stock A-27), different from the amplification products obtained with T. cruzi and T. rangeli. This organism differs from T. cruzi mainly by the size of its trypomastigote forms and kinetoplasts and the absence of infectivity for macrophages and triatomine bugs. It is also morphologically distinct from salivarian trypanosomes reported in Brazil. Isoenzyme analysis at 8 loci demonstrated a very peculiar banding pattern clearly distinct from those of T. rangeli and T. cruzi. We conclude that this isolate is a new Trypanosoma species. The name T. caninum is suggested.

Catabolism of fish furan fatty acids to urofuran acids in the rat.

A mixture of long-chain furan fatty acids was prepared as methyl esters from testes lipids of Northern pike (Esox lucius). Upon feeding these esters to rats, dicarboxylic acids, which still contained the furan structure, were found in the urine. The first phase of a rapid but incomplete catabolism is beta-oxidation of the proximal chain of the furan fatty acids. It proceeds to a distance of three carbon atoms from the ring. omega-Oxidation of the terminal alkyl chain, followed by alpha-oxidation gives rise to a second alkylcarboxyl chain with five carbon atoms or less. The ring methyl substituents of the precursor acids seem to be more resistant to oxidation than the alkyl substituent with three or five carbon atoms. The urinary catabolites from furan fatty acids in the rat are similar to furan acids found in human urine, but only one of the structures occurs in both sources.

The combined dexamethasone-CRH test before and after repetitive transcranial magnetic stimulation (rTMS) in major depression.

BACKGROUND: Hypothalamic-pituitary-adrenocortical (HPA) dysregulation assessed by the combined dexamethasone corticotropin releasing hormone test (DEX/CRH test) has been demonstrated to normalize after successful antidepressant pharmacotherapy. Here, we investigated whether repetitive transcranial magnetic stimulation (rTMS) also leads to a normalization of HPA system activity in depressed patients. METHODS: Thirty-seven medication free patients suffering from a major depressive episode (DSM-IV) underwent a DEX/CRH test before and after 13 daily sessions of left prefrontal rTMS in an open trial. RESULTS: There was an overshoot of CRH-induced cortisol release that was not affected by rTMS treatment. Postdexamethasone cortisol levels prior to CRH challenge decreased in responders after rTMS treatment, whereas no change of CRH-induced adrenocorticotropic hormone (ACTH) and cortisol release in responders or nonresponders was observed. CONCLUSIONS: The persisting HPA system hyperactivity after rTMS suggests a high risk for relapse and therefore argues for an immediate maintenance therapy in patients responding to this treatment.

Histological assessment of nerve lesions caused by epineurial electrode application in rat sciatic nerve.

The left sciatic nerve of 36 rats was exposed and four ring-shaped stainless steel wire electrodes were sutured to the epineurium of each nerve in the same manner as performed clinically for "carousel stimulation" in man. The rats were sacrificed 10 days (Group 1), 3 weeks (Group 2), or 3 months (Group 3) after implantation. The electrodes were excised, the nerves were embedded in Epon, and semithin sections were obtained for histological and planimetric assessment of lesions caused by the epineurially sutured electrodes. The right sciatic nerves served as controls. The total area of neural tissue within the perineurium was determined at three levels: at the site of the electrodes, 8 mm proximal, and 8 mm distal. The area of neural tissue damaged by the surgical procedure was expressed as a percentage of the total area. In Group 1, nine of 12 nerves showed lesions ranging from 0.39% to 25.39% of the total area of neural tissue, in Group 2 eight of 11 sciatic nerves showed lesions ranging from 0.24% to 13.03% of the total area, and in Group 3 five of 12 nerves showed lesions ranging from 0.21% to 4.96% of the total area. The pathologically altered areas in Groups 2 and 3 exhibited distinct signs of nerve fiber regeneration. The reasons for the decrease in damage from Group 1 to Group 3 and the clinical implications of the results for long-term electrical stimulation are discussed.

Applications of the in vitro aryl hydrocarbon hydroxylase induction assay for determining "2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents": pyrolyzed brominated flame retardants.

The pyrolysis of brominated flame retardants FR 300 BA (decabromobiphenyl) ether, FireMaster BP-6 (polybrominated biphenyls), Bromkal 70-5-DE (primarily pentabromodiphenylether), Bromkal 70-DE (primarily penta and tetrabromodiphenylether) and Bromkal G1 (pentabromodiphenylether) resulted in the formation of relatively high levels of polybrominated dibenzofurans (PBDFs) and dibenzo-p-dioxins (PBDDs as determined by gas chromatography-mass spectrometric analysis. The dose response EC50 values for the induction of aryl hydrocarbon hydroxylase (AHH) and ethoxyresorufin O-deethylase (EROD) by the flame retardant pyrolysates was determined in rat hepatoma H-4-II E cells and compared to the relative induction activities of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and the concentrations of "2,3,7,8-TCDD equivalents" were calculated. The range of "2,3,7,8-TCDD equivalents" levels (micrograms/g or ppm) derived from values obtained from the AHH and EROD bioassays for each of the pyrolyzed flame retardant samples was: 174-194, 480-1400, 2140-4680, 6740-8780 and 3920-5260 ppm for FR 300 BA, FireMaster BP-6, Bromkal 70 DE, Bromkal 70-5 DE and Bromkal G1, respectively. The in vivo dose-response effects of 2 pyrolyzed flame retardants were determined in immature male Wistar rats and compared to the dose-response activities of 2,3,7,8-TCDD. The in vivo responses which were measured included hepatic microsomal AHH and EROD induction, body weight loss and thymic atrophy. For the pyrolyzed FireMaster BP-6 and Bromkal 70-5 DE samples, the range of calculated in vivo "2,3,7,8-TCDD equivalents" (ppm in sample) for the 4 in vivo bioassays was 520-1780 ppm and 3860-8960 ppm, respectively. The excellent overlap between the in vivo and in vitro 2,3,7,8-TCDD equivalents for the 2 flame retardant pyrolysate extracts supports the utility of the in vitro induction bioassay for quantitatively determining "2,3,7,8-TCDD equivalents" for mixtures containing toxic halogenated aryl hydrocarbons.

Repetitive transcranial magnetic stimulation (rTMS) in pharmacotherapy-refractory major depression: comparative study of fast, slow and sham rTMS.

In previous studies, fast repetitive transcranial magnetic stimulation (rTMS) with a frequency > 1 Hz demonstrated substantial antidepressant effects compared to sham rTMS. However, it is not clear whether fast rTMS is superior to slow rTMS (frequency < or = 1 Hz) which is safe at therapeutically promising higher intensities. The aim of this double-blind study was to compare the action of fast, slow and sham rTMS. Eighteen patients with pharmacotherapy-resistant major depression were randomized to receive fast (10 Hz), slow (0.3 Hz) or sham rTMS with 250 stimuli/day for 5 successive days. rTMS was applied at 90% motor threshold intensity to the left dorsolateral prefrontal cortex. Scores on the Hamilton Depression Rating Scale (HDRS), but not on the Montgomery-Asberg Depression Rating Scale (MADRS), showed a statistically significant time x group interaction with a reduction of 19% after slow rTMS. However, the effect was clinically marginal and not reflected by self-rating scores. Verbal memory and reaction performance were not impaired after rTMS, and there was even a statistically significant time x group interaction with improvement of verbal memory performance after fast rTMS. In conclusion, this study further supported the safety of rTMS but does not show any clinically meaningful antidepressant efficacy of rTMS at 250 daily stimuli over 5 days in pharmacotherapy-refractory major depression.

Clinical experience with a new recombinant hepatitis-B vaccine in previous non-responders with chronic renal insufficiency.

Patients with renal insufficiency have an increased risk of hepatitis B infection and a high probability to develop a chronic course of this disease. After hepatitis B vaccination they are known to show a low rate of seroconversion. In the present study we assessed the efficacy of a new recombinant pre-S1 and pre-S2 containing hepatitis B vaccine in 17 non-responders (anti-HBs titer 0) and 4 low-responders (anti-HBs titer < or = 5 IU/ml) with chronic renal insufficiency (16 on chronic hemodialysis therapy and 5 without hemodialysis treatment). Seroconversion rate was 65% after the third and 71% after the fourth vaccination. Only minor side effects were seen. These results encourage to use the new vaccine in a larger number of patients with renal insufficiency.

Psychopathological changes preceding motor symptoms in Huntington's disease: a report on four cases.

Neurodegenerative disorders often exhibit "classical" psychiatric symptoms as an initial presentation of the disease. Here we present four patients with different psychopathological abnormalities who were later diagnosed as having Huntington's disease. The range of symptoms covered affective and psychotic symptoms, antisocial behavior, cognitive problems reminiscent of dementia and suicidal idealisation. The pattern of progress of neuronal degeneration may be helpful in explaining the antecedent manifestation of psychiatric symptoms.

Investigation on organic pollutants from a domestic heating system using various solid biofuels.

Various herbaceous biofuels (straw, whole plant cereals and set aside hay) and spruce wood were tested for their potential to form PCDD/F, PCPh, PCBz and PAH during combustion. The trials were conducted in an automatically charged multi-fuel furnace for domestic applications (50 kWth). Both, flue gas and the different ash fractions were analysed. CO-emission results show, that combustion conditions were relatively uniform (mean CO-level: 200 mg/m3 at 13% O2 in flue gas). Likewise, the TOC- and PAH-emissions in the fuel gas remained constantly on a relatively low level. However, for the PCDD/F, PCPh and PCBz increased emissions were detected when herbacious fuels were applied. This may be attributed to their higher chlorine concentration and the high ash content, which is responsible for increased dust emissions during combustion. Similar observations were also found for the PCDD/F-concentrations in the ashes. Combustion chamber ashes usually showed a drastically reduced contamination with highly toxical compounds, compared to the ash fraction from the heat exchanger ash or to chimney soot.

PCDD/F- and PAH-emission from house heating systems.

Combustion experiments were carried out in different house heating systems using various fuels. The emission samples and the chimney soot samples were analyzed for polychlorinated dibenzo-p-dioxins/furans (PCDD/F) and polycyclic-aromatic-hydrocarbons (PAH). The PCDD/F-concentrations in fluegases using untreated wood were in the range of 2-25 pg I-TE/m3. The combustion of paper, cartons, painted wood and wood with 2-5% PVC gave PCDD/F-concentrations from 38 to 952 pg I-TE/m3 and 380 to 2,240 ng I-TE/kg (chimney soot). The PAH-concentrations in flue gas were about 30 times higher using an old technology in relation to a new one.

Polyhalogenated dibenzo-p-dioxins and dibenzofurans in the exhaust air during textile processings.

5 textile finishing processes were investigated for polychlorinated dibenzo-p-dioxins/furans (PCDD/F), polybrominated dibenzo-p-dioxins/furans (PBDD/F) and mixed polychlorinated-brominated dibenzo-p-dioxins/furans (PBCDD/F). For the purpose of these investigations the complete balance--dioxins input from the raw textiles, output over the finished textiles and output through the air path--was done. In the exhaust air only small PXDD/F concentration were detected. The textiles contain before and after the process nearly the same PCDD/F-concentrations. The concentrations of PBCDD/F arises after the process. Only the chimney depositions contain higher PCDD/F-concentrations up to 1,806.1 ng I-TE/kg, PBDD/F-concentrations up to 1,572.6 ng/kg and PBCDD/F-concentrations up to 40,801.4 ng/kg.

Application of a purge and trap TDS-GC/MS procedure for the determination of emissions from flame retarded polymers.

Emissions of volatile organic compounds (VOCs) from different thermoplastic polymers used in electrotechnical applications were investigated using a purge and trap procedure that involved adsorption on Tenax GR. Results were compared to those for an operating TV set monitored in a test chamber. The analyses were in both cases carried out using thermodesorption gas chromatography with mass spectrometric detection (TDS-GC/MS). Substances identified were monomers, volatile additives, or related compounds. Special attention was given to the detection of halogenated compounds. Their origin was studied using reference samples and synthetic standards.

Analysis of flame retarded polymers and recycling materials.

Recycling activities on polymeric materials are increasing and becoming more and more important in recent years. For polymers containing no flame retardants, suitable recycling strategies already exist. In order to investigate the recyclability of flame retarded polymers that contain brominated flame retardants, a number of samples were analysed as received from a recycling company. Following the identification and sorting of the samples according to type of polymers and flame retardants, material recycling was tested for the flame retarded polymers identified to occur most frequently. The reactivity of the flame retardants during the recycling procedure was studied by analysing for brominated dioxins and furans. The results demonstrate that flame retarded polymers can be recycled under certain experimental conditions.

Multichannel stimulation of phrenic nerves by epineural electrodes. Clinical experience and future developments.

Between 1983 and 1992, 23 patients with complete ventilatory insufficiency of differing etiologies were treated with an eight channel implant (Medimplant Inc., Vienna) for fatigue free stimulation of both phrenic nerves. Data for 15 patients with high spinal cord lesions (ages: 9-51 years) are summarized: 1) level of lesion: C0, 3 patients; C1/C2, 4; C2/C3, 8; 2) time between incident and implantation: 3-14 months; 3) diaphragm training: 1-22 months; 4) chronic pacing: 5-83 months; 5) tracheostomy closed: 7 patients; 6) living permanently at home: 13 patients; 7) respiratory rate per minute: 12-17; 8) duration of inspiration: 1.0-1.3 sec; 9) tidal volume: 7-20 ml/kg body weight; 10) volume per minute: 121-198 ml/kg body weight; 11) pH: 7.39-7.42; 12) pCO2: 22.9-38.6 mmHg; 13) pO2: 81.2-104.5 mmHg; and 14) died by December 1992, 4 patients. All currently available implants for phrenic pacing need an external power supply and radio control. The authors have developed and tested the first fully implantable device. Features of this implant include an electronic circuit based on the microcontroller MC68HC705C8; surface mounted technology (SMD); eight channels; constant current source adjustable to 5 mA in 256 steps, impulse duration: 100-1000 musec, stimulation frequency: 1-33 Hz; and minimum lifetime: 3 years. The implant is programmed via bidirectional radio transmission using an IBM compatible computer. The dimensions, including battery, eight electrode connectors, and antenna, are 67 x 48 x 13 mm. The implant weights 58 g. This new device may improve patients' safety and quality of life in the near future.

Effect of stationary guiding vanes on improvement of the washout behind the rotor in centrifugal blood pumps.

In centrifugal pumps, there always exists an area of stagnation between the rear of the rotor and the rear housing wall that promotes thrombus formation around the axle. Some current devices overcome the problem by using holes in the rotor plane, leading to increased hydrodynamic losses and shear stress. In this study, a simple apparatus was developed to overcome this problem. Guiding vanes were fixed to the rear housing wall. These vanes decrease the tangential velocity of the fluid and thus the centrifugal force, leading to an increased secondary flow toward the axle. The effect of such vanes was studied in videographic and ultrasound studies. An increase of washout and mixing between the flow layers could be demonstrated (stay time < 200 msec versus several seconds without vanes). In the first animal experiment using nonoptimized vanes, there was no thrombus at the back plane or the seal, and only a small thrombus at the transition between axle and rotor. Hemolysis was slightly elevated (3.2 mg/dl versus 2.5 mg/dl in control experiments). In conclusion, it is highly likely that this simple system will improve the flow characteristics in centrifugal pumps.

Monitoring set-up for selection of parameters for detection of hypoglycaemia in diabetic patients.

Recurrent severe hypoglycaemia is often an unsolved problem in diabetic patients under intensified insulin treatment. As no reliable long-term stable blood glucose sensor has yet been developed, registration of other body function changes could help to detect severe hypoglycaemia. A measuring system is described, capable on the one hand of recording EEG, heart rate, peripheral pulse, skin temperature, respiratory movements, skin impedance and arterial blood pressure, and capable of registering plasma glucose, counter-regulatory hormones, symptoms and cognitive performance under experimental conditions during hypoglycaemia, on the other. In a clinical study involving both insulin-induced hypoglycaemia in healthy subjects and insulin-dependent diabetic patients, the practical value and the character of changes of the recorded parameters are investigated. Currently insensitivity to hypoglycaemia, impracticability, complexity or susceptibility to artefacts make use of most parameters unsuitable for hypoglycaemia prevention. It is believed, however, that future efforts could result in indirect registration of hypoglycaemia, including a qualified combination of different parameters, individual adaptation in accordance with particular responses of individual patients, together with new measuring and sensor techniques.

Computerized generation of circadian sensor modal days with continuous glucose monitoring for comparison of various insulin regimens based on insulin glargine in type 1 diabetes.

AIM: Our aims were (1) to design and standardize a statistical approach for data reduction in continuous glucose monitoring, allowing comparison of circadian glycemic patterns in therapeutic subcohorts of patients with type 1 diabetes, and (2) to investigate the applicability of this approach for CGMS assessment in clinical study of basal insulin replacement quality with various timings of basal injections (pre-breakfast, dinner, bedtime) of a new insulin analog. METHODS: Prospective randomized three-arm parallel study with switch over after 6 months for another 3 months of free choice injection time point (options pre-breakfast, pre-dinner and bedtime) of the new insulin analog in 16 type 1 diabetic subjects on functional insulin treatment (FIT: basal, prandial and correctional dosages). CGMS was used at the end of each follow up period of a clinical study. Representative daily profiles were off-line computed as "circadian sensor modal days" for each insulin regimen consisting of consecutive means of hourly glucose values. RESULTS: Although the overall quality of glycemic control (HbAIC) for different regimens did not reach statistical differences, CGMS displayed slightly divergent maximal swings in the course of glycemia (p=0.04-0.08) and allowed--with delineated data reduction procedure--a reliable between treatment comparison. CONCLUSION: Off-line computation of "hourly circadian sensor modal days" for data reduction can be effectively used with CGMS for description of circadian glycemic patterns in type 1 diabetes.

Monitoring of FES-induced muscle activity by continuous EMG-recording.

Functional Electrical Stimulation (FES) requires information on the stimulated muscle for adjustment of the stimulation current, avoidance of muscle fatigue during the conditioning period and long term follow-up. Several applications of chronical FES are in clinical practice, but a system for direct registration of muscle activity under FES still does not exist. In six sheep the right Latissimus Dorsi Muscle (LDM) and Thoracodorsal Nerve were exposed. Stimulation electrodes were applied to each nerve and 3 EMG-applied sensing electrodes were placed into each LDM. The LDM tendon was connected to a force transducer. Burst stimulation was applied and the amplitude was increased from 0 to 4 mA in steps from burst to burst. EMG (M-wave) was amplified and recorded continuously via modified instrumentation amplifier, oscilloscope and tape recorder. Isometric muscle tension was recorded using force transducer, A/D interface and PC. Continuous EMG-recording was performed in all cases. Simultaneous recording of muscle tension and EMG revealed a close correlation (IrI=0.95, p < 0.0001) between the muscle strength and amplitude of the M-wave. Continuous recording of the EMG seems to be a reliable method for direct monitoring of the stimulated muscle. Three intramuscular electrodes can provide enough information to monitor FES induced muscle activity.

Minimization of hemolysis in centrifugal blood pumps: influence of different geometries.

Centrifugal blood pumps are of substantial importance for intraoperative extracorporeal circulation and for temporary cardiac assist. Their development and improvement raises many specific questions, especially on mechanical blood properties, flow distribution, and the resulting biocompatibility. In this comprehensive study the influence of various pump geometries on blood trauma was investigated. For this purpose analytical calculations, hydrodynamic performance, numerical simulation, in vitro hemolysis tests and in vivo experiments were used. The gap between rotor and housing was found to be crucial showing a distinct minimum of hemolysis at a gap of 1.5 mm (in vitro increase of plasma free hemoglobin per 100 ml plasma an hour: delta fHb/hour = 2.4 +/- 0.83 mg%/h at 1.5 mm versus 12 +/- 2.2 mg%/h at 2.5 mm; p < 0.05). Housing diameter and shape of the vanes were of less importance for blood traumatization (d = 60 mm: delta fHb/hour = 6.36 +/- 1.8 mg%/h; d = 70 mm: fHb = 7.1 +/- 1.9 mg%/h; straight radial vanes: 5.2 +/- 1.8 mg%/h; straight inclined vanes: 6.8 +/- 1.2 mg%/h; flexed vanes: 6.1 +/- 2.0 mg%/h). Three animal experiments confirmed the optimization of geometry, with a mean fHb of 2.5 to 3.2 mg% in steady state. Hydrodynamic efficiency revealed to be a necessary, but not a sufficient and sensitive criterion for hemolysis minimization (e.g. changes of eta < 10% for changes of fHb > 500%). Numerical simulation gives an improved insight in flow distribution, but can not yet be applied for quantification of blood trauma.(ABSTRACT TRUNCATED AT 250 WORDS)

Laryngeal pacemaker: activity of the posterior cricoarytenoid muscle (PCM) and the diaphragm during respiration in sheep.

Electromyograms (EMG) of the PCM and the diaphragm were evaluated in 6 sedated female sheep. Corresponding pneumatograms were recorded simultaneously by means of thermocontrolled respiration flowmeter. Evidence was obtained on considerable intra- and inter-individual differences in the duration of respiratory cycles as well as PCM- and diaphragmatic activity. Most of the evaluated respiratory periods showed either a phase coincidence between the PCM and the diaphragm, or a leading edge of about 40-80 ms of the posticus muscle. Due to this minimal phase shift, the diaphragmatic myogram seems to be a valuable trigger for an external PCM stimulation unit in bilateral recurrent nerve palsy.